IRF3 Antibody (YA334)

IRF3 Antibody (YA334) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to IRF3.

IRF3, a key transcriptional regulator, plays a critical role in orchestrating type I interferon (IFN)-dependent immune responses against DNA and RNA viruses. It regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes by binding to an interferon-stimulated response element in their promoters, exhibiting greater potency in activating the IFN-beta gene. In uninfected cells, IRF3 remains in an inactive form in the cytoplasm, but upon viral infection, recognition of double-stranded RNA or toll-like receptor signaling leads to its phosphorylation by IKBKE and TBK1 kinases. This phosphorylation induces a conformational change, promoting dimerization, nuclear localization, and association with CREB binding protein to form dsRNA-activated factor 1 (DRAF1). This complex activates the transcription of type I IFN and IFN-stimulated genes, driving both early and late phases of gene induction. In the absence of viral infection, IRF3 is maintained as a monomer in an autoinhibited state, and its liberation for DNA binding and dimerization, along with nuclear localization, follows phosphorylation by TBK1 and IKBKE. This phosphorylation occurs in response to viral RNA, cytosolic DNA, or bacterial lipopolysaccharide, initiating a cascade involving innate adapter proteins (MAVS, STING1, or TICAM1) and culminating in the activation of IRF3, which subsequently induces IFNs in the nucleus.

Free

Q14653

Predicted band size: 47 kDa; Observed band size: 55 kDa

Protein A affinity purified.

Cytoplasm, Nucleus.

Non-conjugated

Unmodified

AB_3102577

Immunology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:5,000 ;ICC/IF: 1:50-1:200 ;IHC-P: 1:50-1:200 ;FC: 1:50-1:100

• 
  Western blot analysis of extracts from Hela (lane 2(20μg) , Jurkat (lane 3(20μg), A549 (lane 4(20μg) and HEK293(lane 5(20μg) using IRF3(HY-P80504) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling IRF3 with IRF3 Antibody (HY-P80504)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with IRF3 Antibody (HY-P80504) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using IRF3 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80504, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using IRF3 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80504, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Flow cytometric analysis of 1X10^6 Jurkat cells labeling IRF3 Antibody (HY-P80504, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/1000 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

WB, ICC/IF, IHC-P, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human IRF3.AA range:71-120.